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Molecular evolution of Pr1 proteases depicts ongoing diversification in Metarhizium spp

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Abstract

The Pr1 family of serine endopeptidases plays an important role in pathogenicity and virulence of entomopathogens such as Metarhizium anisopliae (Ascomycota: Hypocreales). These virulence factors allow for the penetration of the host cuticle, a vital step in the infective process of this fungus, which possesses 11 Pr1 isoforms (Pr1A through Pr1K). The family is divided into two classes with Class II (proteinase K-like) comprising 10 isoforms further split into three subfamilies. It is believed that these isoforms act synergistically and with other virulence factors, allowing pathogenicity to multiple hosts. As virulence coevolves through reciprocal selection with hosts, positive selection may lead to the evolution of new protease families or isoforms of extant ones that can withstand host defenses. This work tests this hypothesis in Class II Pr1 proteins, focusing on M. anisopliae, employing different methods for phylogenetic inference in amino acid and nucleotide datasets in multiple arrangements for Metarhizium spp. and related species. Phylogenies depict groups that match the taxonomy of their respective organisms with high statistical support, with minor discrepancies. Positively selected sites were identified in six out of ten Pr1 isoforms, most of them located in the proteolytic domain and spatially close to the catalytic residues. Moreover, there was evidence of functional divergence in the majority of pairwise comparisons. These results imply the existence of differential selective pressure acting on Pr1 proteins and a potential new isoform, likely affecting host specificities, virulence, or even adapting the organism to different host-independent lifestyles.

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Acknowledgements

The authors would like to thank BV, CLF, CLM, JFRB, GPP, MT, RKC, RLB, and NSO for insightful discussions, suggestions and overall support. This work was supported by the following Brazilian agencies: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) [Grant: Biocomputacional 23038.010041/2013-13, Rede Avançada em Biologia Computacional (RABICÓ)], Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) [Grant: Universal 2014 458160/2014-8].

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  1. Augusto Schrank and Claudia Elizabeth Thompson share senior authorship.

Authors and Affiliations

  1. Rede Avançada em Biologia Computacional (RABICÓ), Petrópolis, RJ, Brazil

    Fabio Carrer Andreis, Augusto Schrank & Claudia Elizabeth Thompson

  2. Centro de Biotecnologia, Programa de Pós-Graduação em Biologia Celular e Molecular (PPGBCM), Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Fabio Carrer Andreis, Augusto Schrank & Claudia Elizabeth Thompson

  3. Departamento de Farmacociências, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, Brazil

    Claudia Elizabeth Thompson

  4. Programa de Pós-Graduação em Ciências da Saúde, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, Brazil

    Claudia Elizabeth Thompson

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  1. Fabio Carrer Andreis
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Correspondence to Claudia Elizabeth Thompson.

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Andreis, F.C., Schrank, A. & Thompson, C.E. Molecular evolution of Pr1 proteases depicts ongoing diversification in Metarhizium spp. Mol Genet Genomics 294, 901–917 (2019). https://doi.org/10.1007/s00438-019-01546-y

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